Enhanced Production of Ganoderic Acids and Cytotoxicity of Ganoderma lucidum Using Solid-Medium Culture

Submerged cultures of Ganoderma lucidum are used to produce fungal mycelium, which is used as a functional food and in the production of various triterpenoids, including ganoderic acids (GAs). Specific culture approaches that produce fungal mycelium with high levels of GAs and good biological activity are critical in the functional food industry. In this study, a solid-medium culture approach to producing mycelium was compared to the submerged culture system. Production of GAs, biomass, intracellular polysaccharides, and cytotoxicity of the cultured mycelium were compared as between solid and submerged culture. Growing G. lucidum strains on solid potato dextrose agar medium increased biomass, the production of ganoderic acid 24 (lanosta-7,9(11), 24-trien-3α-o1-26-oic acid), GAs, and total intracellular polysaccharides as compared to fungi grown in submerged culture. Triterpenoid-enriched methanol extracts of mycelium from solid-medium culture showed higher cytotoxicity than those from submerged culture. The IC(50) values of methanol extracts from solid-medium culture were 11.5, 8.6, and 9.9 times less than submerged culture on human lung cancer cells CH27, melanoma cells M21, and oral cancer cells HSC-3 respectively. The squalene synthase and lanosterol synthase coding genes had higher expression on the culture of solid potato dextrose medium. This is the first report that solid-medium culture is able to increase GA production significantly as compared to submerged culture and, in the process, produces much higher biological activity. This indicates that it may be possible to enhance the production of GAs by implementing mycelium culture on solid medium.     

细胞外信号调节激酶1/2信号通路在慢性哮喘模型大鼠支气管平滑肌细胞迁移功能变化中的调控作用

本研究旨在探讨细胞外信号调节激酶1／2(extracellular signal-regulated kinase,ERK1/2)信号通路在慢性哮喘模型大鼠支气管平滑肌细胞(bronchial smooth muscle cells,BSMCs)迁移能力改变中的调控作用。应用卵清蛋白致敏和雾化方法制备大鼠慢性哮喘模型,体外培养大鼠BSMCs,采用免疫荧光细胞化学、Western blot和RT-PCR方法检测ERK1／2信号通路的表达,分别用平面迁移实验和跨膜迁移实验来评价BSMCs的活动和趋向迁移能力,并比较用和不用ERK1／2信号通路干预剂的差异。Western blot结果显示慢性哮喘模型大鼠BSMCs中总ERK1／2(9．13±0．87)较对照组(4．68±0．59)明显增加,磷酸化ERK1／2(p-ERK1/2)占总ERK1／2的比值(0．55±0．05)较对照组(0．48±0．04)显著提高(n=10,P<0．01)。慢性哮喘组ERK1和ERK2 mRNA的表达(1．83±0．24和1．07±0．11)较对照组(0．58±0．14和0．51±0．12)明显增高(n=10,P<0．01)。在平面迁移实验中,慢性哮喘大鼠BSMCs的迁移最远距离是对照组的(2．9±0．1)倍,在ERK1／2激动剂表皮生长因子(epidermal growth factor, EGF)刺激下增加到(5．0±0．2)倍,而在30μmol／L PD98059的作用后下降到(1．7±0．2)倍。正常对照大鼠BSMCs平面迁移能力对PD98059的反应较慢性哮喘组弱,仅在100μmol／L PD98059的作用下下降到(0．8±0．1)倍。跨膜迁移实验中,慢性哮喘大鼠BSMCs的跨膜迁移细胞是对照组的(1．9±0．1)倍,在EGF刺激下增加到(3．1±0．2)倍,而在30μmol/L PD98059作用后下降到(1.45±0.2)倍。这些结果表明慢性哮喘模型大鼠BSMCs的迁移能力明显增强,ERK1／2信号通路在该功能变化的调控中可能发挥了重要作用。     

Development and function of central cell in angiosperm female gametophyte

The central cell characterizes the angiosperm female gametophyte (embryo sac or megagametophyte) in that it directly participates in “double fertilization” to initiate endosperm development, a feature distinguishing angiosperm from all other plant taxa. Polygonum‐type central cell is a binucleate cell that, upon fertilization with one of the two sperm cells, forms triploid endosperm to nourish embryo development. Although the formation and the structure of central cell have well been elucidated, the molecular mechanisms for its specification and development remain largely unknown. The central cell plays a critical role in pollen tube guidance during pollination and in endosperm initiation after fertilization. Recently, a group of mutants affecting specific steps of central cell development and function have been identified, providing some clues in understanding these questions. This review summarizes our current knowledge about central cell development and function, and presents overview about hypotheses for its evolution. genesis 48:466–478, 2010. © 2010 Wiley‐Liss, Inc.     

奥扎格雷钠联合依达拉奉治疗急性脑梗死的疗效观察

目的:观察奥扎格雷钠联合依达拉奉治疗急性脑梗死的临床疗效。方法:将160例急性脑梗死患者随机分为治疗组和对照组,每组各80例,治疗组采用注射用奥扎格雷钠联合依达拉奉治疗,对照组仅采用注射用奥扎格雷钠治疗,观察两组患者入院及治疗14d后临床疗效,神经功能缺损评分变化并进行对比分析。结果:治疗组的显效率为85.9%(67/78),对照组显效率为70.9%(56/79),两组显效率有显著性差异(x2=5.12,P=0.022),两组患者治疗14d后,神经功能的恢复情况与治疗前相比均有显著性差异(P<0.05),且治疗组与对照组治疗后比较有显著性差异,具有统计学意义(P<0.05)。结论:奥扎格雷钠联合依达拉奉治疗急性脑梗死较单独使用奥扎格雷钠疗效显著,是治疗急性脑梗死的有效方法。     

Water transport of native and exotic tree species in relation to xylem anatomical characteristics in low subtropical China

Aims Exotic fast-growing tree species have been commonly planted as pioneer species to facilitate ecological restoration in South China. Their growth and resource utilization behavior related to intrinsic physiology and structural properties have profound influences on forest ecosystem. However, the contrastive research focusing on water utilization features along with xylem anatomical properties between native and exotic species is scarce in South China. The objective of this study is to investigate the sapwood anatomical characteristics and water utilization conditions of native and exotic fast-growing species, and to elucidate the relationship between sap-flux density and conduit features.Methods We measured sap-flux density, conduit length, diameter and density of four native species (Schima superba, Michelia macclurei, Castanopsis hystrix and Castanopsis fissa) and four exotic species (Eucalyptus citriodora, Eucalyptus urophylla × grandis, Acacia auriculaeformis and Acacia mangium). Sap flux density was measured based on the Granier's thermal dissipation probe method. The whole-tree water transport was quantified by multiplying sap-flux density by sapwood area. The measurements of conduit characteristics were conducted by using segregation and slice method.Important findings Sapwood area increased with the growing diameter at breast height (DBH) as a power function. Native species had a larger water-conducting tissue area than exotic species at the same DBH value when trees grew to a size with a certain value of DBH. The conduit diameter of exotic species was significantly larger than that of native species. Conversely, native species, such as S. superba and M. macclurei, had longer conduit length and higher conduit density than other tree species. Based on a physiological interpretation of the measured conduit characteristics, native tree species developed a safe water transport system while exotic fast-growing tree species come into being an efficient system instead. Water transport increased with the growing DBH as a power function, and the exponent for native species (1.60) was higher than that for exotic species (1.22). Under the combined impact of sap-flux density and sapwood area, native species presented a larger water transport at a larger DBH value, indicating that growth advantage of exotic fast-growing species might weaken as DBH increased.     

新疆额尔齐斯河流域冷水鱼肠道耐低温乳酸菌的分离筛选及其遗传差异

[目的]从新疆阿勒泰地区额尔齐斯河流域冷水鱼肠道中分离耐低温的乳酸菌,挖掘乳酸菌的物种和遗传多样性,为低温乳酸菌生物技术研发提供优良菌种.[方法]利用MRS、Elliker两种不同培养基从9种冷水鱼肠道中,分离筛选出耐低温菌,测定细菌最适生长温度并进行常规生理生化实验.根据16S rRNA基因序列初步确定耐低温菌的系统进化地位,并采用BOX、(GTG)5、ERIC三种不同的引物进行rep-PCR,对16S rRNA基因高度同源性的菌株进一步区分.[结果]分离得到78株耐冷菌,最终确定有24株为耐低温乳酸菌,菌株的最适生长温度在15-24℃之间.系统发育分析结果表明:24株耐低温乳酸菌隶属于6个属,其中肉杆菌属(Carnobacterium)3株,乳球菌属(Lactococcus)9株,肠球菌属(Enterococcus)7株,环丝菌属(Brochothrix)1株,魏斯氏菌属(Weissella)2株,链球菌属(Streptococcus)2株.指纹图谱聚类分析树状图显示乳球菌属(Lactococcus)和肠球菌属(Enterococcus)存在种及菌株水平上的遗传差异,前者包括4个种,后者2个种.[结论]新疆额尔齐斯河流域冷水鱼肠道中分离的耐低温乳酸菌以球菌为主,没分离到常见的乳杆菌,需进一步完善分离培养的方法,深入挖掘和研究其物种多样性和遗传多样性.     

甜菜夜蛾半胱天冬酶基因的克隆及对细胞凋亡诱导剂和病原微生物感染的表达响应

[目的]本研究旨在明确甜菜夜蛾Spodoptera exigua半胱天冬酶(caspase)在细胞凋亡诱导剂诱导和病原微生物胁迫下的表达模式,为丰富鳞翅目昆虫细胞凋亡机制研究奠定基础.[方法]利用RT-PCR技术从甜菜夜蛾3龄幼虫体内扩增两个半胱天冬酶基因(SeCasp-3和SeCasp-4)编码区的全长;利用qPCR...     

Variations of cuticular wax in mulberry trees and their effects on gas exchange and post-harvest water loss

Though mulberry (Morus alba) tree shows great adaptations to various climate conditions, their leaf water status and photosynthesis are sensitive to climate changes. In the current study, seven widely planted mulberry cultivars in Chongqing, Southwest China, were selected to analyze leaf cuticular wax characteristics, gas exchange index, post-harvest leaf water status and their relationships, aiming to provide new theory in screening high resistant mulberry cultivars. Mulberry trees formed rounded cap-type idioblasts on the adaxial leaf surface. Film-like waxes and granule-type wax crystals covered leaf surfaces, varying in crystal density among cultivars. The stomatal aperture on the abaxial surface of cultivars with high wax amount was smaller than that of cultivars with low wax amount. The amount of total wax was negatively correlated with the net photosynthetic rate (P _N), transpiration rate (E) and stomatal conductance (g _s) and positively correlated with the moisture retention capacity. It suggested that both cuticular wax and stomatal factor might be involved in regulating water loss in mulberry leaves under field conditions. The variability in moisture retention capacity and cuticular wax characteristics might be important in evaluating and screening mulberry cultivars for increasing silk quality and silkworm productivity.     

Antimicrobial Resistance and Molecular Epidemiology of Escherichia coli Causing Bloodstream Infections in Three Hospitals in Shanghai,China

Escherichia coli (E. coli) is one of the most frequent and lethal causes of bloodstream infections (BSIs). We carried out a retrospective multicenter study on antimicrobial resistance and phylogenetic background of clinical E. coli isolates recovered from bloodstream in three hospitals in Shanghai. E. coli isolates causing BSIs were consecutively collected between Sept 2013 and Sept 2014. Ninety isolates randomly selected (30 from each hospital) were enrolled in the study. Antimicrobial susceptibility testing was performed by disk diffusion. PCR was used to detect antimicrobial resistance genes coding for β-lactamases (TEM, CTX-M, OXA, etc.), carbapenemases (IMP, VIM, KPC, NDM-1 and OXA-48), and phylogenetic groups. eBURST was applied for analysis of multi-locus sequence typing (MLST). The resistance rates for penicillins, second-generation cephalosporins, fluoroquinolone and tetracyclines were high (>60%). Sixty-one of the 90 (67.8%) strains enrolled produced ESBLs and no carbapenemases were found. Molecular analysis showed that CTX-M-15 (25/61), CTX-M-14 (18/61) and CTX-M-55 (9/61) were the most common ESBLs. Phylogenetic group B2 predominated (43.3%) and exhibited the highest rates of ESBLs production. ST131 (20/90) was the most common sequence type and almost assigned to phylogenetic group B2 (19/20). The following sequence types were ST405 (8/90) and ST69 (5/90). Among 61 ESBL-producers isolates, B2 (26, 42.6%) and ST131 (18, 29.5%) were also the most common phylogenetic group and sequence type. Genetic diversity showed no evidence suggesting a spread of these antimicrobial resistant isolates in the three hospitals. In order to provide more comprehensive and reliable epidemiological information for preventing further dissemination, well-designed and continuous surveillance with more hospitals participating was important.